Binding of galectin-3 by low molecular weight pectin

ABSTRACT

Administration of low molecular weight (10,000-20,000 Daltons, or lower) pectins, particularly modified citrus pectins (MCP), like PectaSol-C reduces galectins-3 levels in vivo. Reduction of galectin-3 levels by MCP inhibits inflammation, inhibits fibrosis formation in organs and tissues, and inhibits cancer formation, progression, transformation and metastases. The reduction in circulating, serum and cellular galectin-3, inherently resulting from the administration of MCP, provides benefit over a spectrum of biological conditions, as evidenced by in vivo trials.

PRIORITY DATA AND INCORPORATION BY REFERENCE

This application is a continuation-in-part of U.S. patent applicationSer. No. 11/485,955, filed Jul. 14, 2006, pending, which claims thebenefit under 35 U.S.C. § 119(e) of U.S. Provisional Patent ApplicationSer. No. 60/711,415, filed on Aug. 26, 2005, both of which areincorporated by reference herein in their entirety. This applicationalso claims benefit of priority to U.S. patent application Ser. No.12/984,843, filed Jan. 5, 2011, pending, and U.S. Provisional PatentApplication Ser. No. 61/447,138 filed Feb. 28, 2011 both of which areincorporated by reference in their entirety.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to improvement of mammalian, includinghuman, biological conditions impacted by, or mediated by, galectin-3.Galectin-3 is a member of the lectin family, of which at least fourteen(14) mammalian galectins have been identified. Galectin-3 isapproximately 30 kDa and, like all galectins, contains acarbohydrate-recognition-binding domain (CRD) of about one hundredthirty (130) amino acids that enable the specific binding ofβ-galactosides. Galectin-3 is encoded by a single gene, LGALS3, locatedon chromosome 14, locus q21-q22. It is expressed in the nucleus,cytoplasm, mitochondrion, cell surface, and extracellular space, and cancirculate in the blood stream. This protein has been shown to beinvolved in a large number of biological processes, including celladhesion, cell migration, cell invasion, cell activation andchemoattraction, cell growth and differentiation, cell cycle, andapoptosis. Given galectin-3's broad biological functionality, it hasbeen demonstrated to be involved in a large number of disease states ormedical implications. Studies have also shown that the expression ofgalectin-3 is implicated in a variety of processes associated with heartfailure, including myofibroblast proliferation, fibrogenesis, tissuerepair, inflammation, and ventricular and tissue remodeling. Elevatedlevels of galectin-3 in the blood have been found to be significantlyassociated with higher risk of death in both acute decompensated heartfailure and chronic heart failure populations, as well as constituting abiomarker of cancer progression to a metastatic stage.

Background of the Invention

In 2006 U.S. patent application Ser. No. 11/485,955 was filed, a utilitypatent application claiming priority from a 2005 provisionalapplication. This patent application, making reference to earlierdisclosures of administration of modified alginates and pectins, such asthose in U.S. Pat. Nos. 6,274,566 and 6,462,029, disclosed for the firsttime the utility of using very specific low molecular weight pectins,such as PectaSol-C MCP available from EcoNugenics of Santa Rosa, Calif.This modified citrus pectin (MCP) and similar very low molecular weightpectins (molecular weight of 20,000 Daltons or less, preferably about10,000 Daltons) is shown in U.S. patent application Ser. No. 11/485,955to be effective in stimulating a variety of immune responses in mammals.

In the years since the filing which first set forth the administrationof these low molecular weight modified citrus pectins, and similarpectins, research has demonstrated that at least one mode of action ofMCP is the binding of galectin-3 molecules. This binding, an inherentfeature of the inventions disclosed in U.S. patent application Ser. No.11/485,955, is also a central mode of action in the later-filed U.S.patent application Ser. No. 12/984,843, filed Jan. 5, 2011 directed tothe inhibition of certain cancers. In fact, it is now clear thatadministering PectaSol-C MCP or other low molecular weight pectins atthe dosage levels of 5-1,500 mg/kg of body weight per day, with apreferred range of 10 mg/kg/day to 1,000 mg/kg/day inherently bindsgalectin-3 molecules in mammals in need of same in a variety ofbiological systems, providing therapeutic benefit against many of thedisease conditions mentioned.

Thus, the activity of galectin-3 in aggravating or promoting cancer, aswell as the ability of a cancer to metastasize, is widely commented onin the literature following the 2006 disclosure of the effectiveness oflow molecular weight pectins like PectaSol-C in promoting immunesystems. These literature findings stress repeatedly the importance ofbinding or reducing the circulating concentration or titer ofgalectin-3, and/or inactivating glaectin-3 through galectin-3 binderssuch as PectaSol-C. See, for example, Wang et al, Cell Death andDisease, 1-10 (2010) (galectin-3 inhibition promotes treatment) and Yuet al, J. Biol. Chemistry, Vol. 282, 1, pp. 773-781 (2007) establishingthat galectin-3 interactions may enhance formation of cancer ortransformation of metastatic cancer.

Similar reports link acceleration of cancer formation and transformationto circulating galectin-3 concentrations, and suggest that reducinggalectin-3 circulating concentrations, reducing its free expression orotherwise reducing available galectin-3 or galectin-3 interactionsimprove cancer prognosis. Zhao et al, Cancer Res. 69, 6799-6806 (2009),Zhao et al, Molecular Cancer 9, 154, 1-12 (2010) and Wang et al, Am. Jof Pathology, 174, 4, 1515-1523 (2009) wherein siRNA-induced reductionof galectin-3 is shown to slow the course of prostate cancer. Clearly,there is substantial literature that supports the conclusion thatreducing circulating galectin-3, either by blocking its expression, orby binding it, as inherently disclosed in the 2006 filing of U.S. patentapplication Ser. No. 11/485,955, is important in controlling cancer.

Circulating galectin-3 is implicated in a wide variety of biologicalconditions, however. Cardiac fibrosis is gaining significant attentionas a complicating risk factor in cardiac disease, and in particular,chronic heart failure (CHF). Lok et al. Clin. Res. Cardiol, 99, 323-328(2010). DeFillipi et al, U.S. Cardiology, 7,1, 3-6 (2010) clearlyindicate that circulating galectin-3 is an important factor in fibrosisof many organs and organ systems, and that reducing circulatinggalectin-3 may have an important role in remediating cardiac injury andprogression to heart failure (HF). Similarly, Psarras et al, Eur. HeartJ., Apr. 26, 2011 demonstrate that reduction in galectin-3 levels in themyocardium may reduce fibrosis in the heart and improve outlook. De Boeret al, Ann. Med., 43,1, 60-68 (2011) identify galectin-3 as a keyindicator in cardiac health. Shash et al, Eur J. Heart Fail., 12,8,826-32 (2011) identify galectin-3 levels as a key agent in heart failurethrough fibrosis. De Boer et al. Eur. J. Heart Fail., 11, 9, 811-817(2009) link an increase in galectin-3 expression and presence toheightened fibrosis, and heart failure. The same article linksgalectin-3 to inflammation. Inflammation is the hallmark ofarteriosclerosis and therefore galectin-3 levels also contribute tocoronary artery disease, peripheral artery disease, strokes, andvascular dementia.

Fibrosis and inflammation, both mediated to some degree by galectin-3(cellular or circulating) are implicated in a variety of conditions ofthe mammalian body, not just cardiac injury and heart failure. Thebinding of galectin-3 achieved by administration of low molecular weightpectins (at least, as reflected in U.S. patent application Ser. No.11/485,955 10,000-20,000 Daltons molecular weight such as PectaSol MCP)is effective in reducing trauma due to kidney injury. Kolatsi-Jannou etal. PlusOne. 6, 4, e18683 (2011). The reduction in circulatinggalectin-3 levels is also indicated to reduce inflammation associatedwith type 2 diabetics, and similar metabolic diseases. Weigert et al, J.Endocrinol. Metab. 95, 3,1404-1411(2010). Thus, high levels ofgalectin-3 have been linked to thyroid cancer, Sethi et al. J. Exp.Ther. Oncol., 8, 4,341-52 (2010) and reduction of galectin-3 expressionand circulation may delay or reduce tumor cell transformation. Chiu etal, Am J. Pathol. 176, 5, 2067-81 (2010).

As noted, galectin-3 is implicated in a wide variety of biologicalconditions, and a reduction in galectin-3 activity, such as that whichcan be achieved by galectin-3 binding with PectaSol-C MCP and similarlow molecular weight pectins may be of value in treating gastriculcerative conditions. Srikanta, Biochimie, 92, 2, 194-203 (2010). Kimet al, Gastroenterology 138, 1035-45 (2010) indicate that reducinggalectin-3 levels ma) be of therapeutic value in reducing gastric cancerprogression. By the same methodology, reducing galectin-3 levelssensitizes gastric cancer cells to conventional chemotherapeutic agents.Cheong et al. Cancer Sci., 101, 1, 94-102 (2010). Galectin-3 isimplicated in a wide variety of gastrointestinal conditions. Reducinggalectin-3, by binding for example, may reduce inflammation in the gutmucosa making MCP an important agent for treatment of ulcerativecolitis, non-specific colitis and ileitis, Crohn's disease, Celiacdisease, and gluten sensitivity. Fowler et al, Cell Microbiol.,81,1,44-54 (2006).

Biliary artesia, a liver disease, is associated with extensive fibrosisof the liver linked with elevated galectin-3 levels. Honsawek et al,Eur. J. Pediatr. Surg., April, 2011. Reduction of galectin-3 levelsresulted in a general improvement in hepatic health, including reducinginflammation, hepatocyte injury and fibrosis. Federici et al. J.Heptal., 54, 5, 975-83 (2011). See also, Liu et al. World J.Gastroenterol. 14,48, 7386-91 (2008) which reported, followingApplicant's teaching in 2005 and 2006 to administer low molecular weightMCP, that MCP inhibited liver metastases of colon cancer and reducedgalectin-3 concentrations. MCP may be used for prevention of liverinflammation, liver fibrosis and liver cirrhosis as well as post-diseaseliver damage, including the various viral hepatitis disease (B, C, andothers) and may be used as well in the treatment of parasitic andchemical hepatitis, chemical liver damage, and others.

SUMMARY OF THE INVENTION

The invention lies in the recognition that the method described in U.S.patent application Ser. No. 11/485,955 of administering low molecularweight pectins, such as modified citrus pectin, having a molecularweight of between 10,000-20,000 Daltons, in a therapeutically effectivedosage, has implications far beyond stimulating the immune system.Because this administration, particularly oral or intravenous,inherently binds cellular, serum and circulating galectin-3, it isimplicated in a wide variety of biological conditions and may beeffective in ameliorating a wide variety of disease states andconditions.

Further improvement can be obtained in selecting the MCP to have amolecular weight of between 3,000-13,000 Daltons that is mostly linearhomogalacturonan with fewer than ten percent (10%) esterification, andmaintaining a percentage of rhamnogalacturonan-1, II in the MCP toapproximately ten percent (10%), while reducing the amount of monogalacturonic acid to under ten percent (10%). The reduction in molecularweight promotes bioavailability (absorption into the circulatory systemfrom the gastrointestinal tract), the degree of esterification (bulkyside groups which contribute to cross bridging between pectin fibers)decreased below ten percent (10%) in the MCP contributes to thegalectin-3 binding by fostering open fibers, allowing increasedaccessibility to binding galectin-3, while the reduction inmono-galacturonic acid (representative of inactive total breakdown ofthe pectin fiber into its major single subunit) increases that amount ofMCP available for effectively binding galcetin-3. Therapeutic propertiesin vivo are further improved by selecting a modified citrus pectin withapproximately ten percent (10%) rhamnogalcturonan-I, II content. Pecticrhamnogalacturonan-1 has been shown to induce apoptosis in melanomacells by interacting with Gal-3. Rhamnogalacturonan-II has demonstrateda strong binding potential for heavy metal chelation. The modifiedcitrus pectin, prepared by enzymatic degradation, results in shorterchain molecules of low esterification with enhanced bio-availability andbinding potential to galectin-3.

This modified citrus pectin can be used to improve or enhance a mammal'simmune reaction, as disclosed in U.S. patent application Ser. No.11/485,955. The power of binding galectin-3, a ubiquitous molecule,without eliminating it from circulation, is powerful beyond this limitedindication, however. Galectin-3 is implicated, as noted, in a widevariety of medical conditions. The inhibition of the growth andtransformation of various cancers, the inhibition of fibrosis in avariety of organs and organ systems, the reduction in inflammationassociated with galectin-3, all combine to provide a powerful method oftreatment in a wide variety of situations. The issue is more of patientselection than treatment modification. In 2006, Applicant taught thatMCP should be administered in a range of low molecular weight pectins atthe dosage levels of 5-1,500 mg/kg of body weight per day, with apreferred range of 10 mg/kg/day to 1,000 mg/kg/day to achieve thedesired results. This range inherently achieves binding or inactivationof cellular, serum and circulating MCP. Particularly for conditionscharacterized at least in part by fibrosis, a reduced preferred range of10 mg/kg/day-750 mg/kg/day may provide even better effects.

PectaSol-C is only one modified pectin useful in this invention. Ingeneral, pectins of reduced molecular weight (10,000 to 20,000 Daltonsor lower) having a lower percentage of mono galacturonic acid and havingapproximately ten percent (10%) rhamnogalcturonan-I, II all exhibit ahigher rate of forming complexes with the multifunctionalcarbohydrate-binding protein, galectin-3, and thus reducing theeffective level of galectin-3 to which a body, injured organs andthreatened cells are exposed. In addition to the available commercialsources of low molecular weight pectins, their preparation is discussedin detail, Pienta et al, J. Nat'l. Cancer Inst., 87, 348-352 (1995). Inan alternative embodiment, low molecular weight pectin can also besynthesized. Specific molecular structures such as, for example,polygalacturonic acid, side branches and neutral sugars in the desiredrange as specified above can be synthesized to create a more consistent,accurate and highly reproducible molecular weight. By synthesizing MCP,an optimal structure within the molecular weight range of 10-20 KD, andmore preferably 3-13 KD can be produced. Whether derived from reducednatural starting materials, or synthesized, administration of theselower molecular weight pectins, 20,000-10,000 Daltons, or even lower,13,000-3,000 Daltons, is a preferred method of practicing the invention.

DETAILED DESCRIPTION OF THE INVENTION

The binding of galectin-3 by MCP is an event that will aid medicalconditions over a wide variety of indications. These include cancer,inflammation and fibrosis, heart disease, kidney damage, liver damage,bladder disease, thyroid disease, pulmonary disease, immune response,stroke, persistent acute inflammation due to non-degradable pathogens,persistent foreign bodies, or autoimmune reactions, hypersensitivitiesand allergies, pesticides, environmental toxins, and heavy metals, aswell as heterogenic conditions such as radiation (examples being medicalprocedures such as various radiation therapies, exposure to ionizingradiation, nuclear radiation, cosmic radiation, electromagneticradiation) and chemotherapy damage, and post radiation and chemotherapyinduced inflammation and fibrosis. As noted above, elevated galectin-3levels are associated with (1) growth, transformation and metastaticmigration of cancer cells across a wide variety of cancers, includingliver, breast, prostate, colon, thyroid, gallbladder, nasopharyngeal,lymphocytic leukemia, melanoma and lung cancers among others, as well asreducing sensitivity in these cancers to conventional antineoplasticagents; (2) development and extension of fibroses beyond normal andhealthy levels, in situations associated with cardiovascular disease andheart failure, in tissue injury including brain, lungs, renal, hepatic,heart and gastroenterologic situations as well as tissue damage due toradiation and chemotherapy exposure and persistent acute inflammationdue to non-degradable pathogens, persistent foreign bodies, orautoimmune reactions, hypersensitivities and allergies; and (3) ininflammation that may be associated with disease or organ failure modes,including diabetes (1 and II), heart disease and dysfunction,atherosclerosis, asthma (bronchial inflammation), ulcers, intestinalinflammation in the bowels (inflammatory bowel diseases), hepaticinflammation associated with both alcohol and non-alcohol relatedcirrhosis and inflammation, liver infections such as viral hepatitisamong others. Other indications associated with inflammation andsusceptible to treatment by administration of MCP includeparasite-induced conditions, such as trypanosomiasis, cerebral malaria,and inflammation and resistance to various infections includingParacoccidiosis brasilensis (fungal infection), schistosomiasis,granulatomatous bronchopneumonia, and in inflammation associated witharthritis and other diseases of the skeletomuscular and skin systems,including inflammation and fibrosis related conditions such as psoriasisand aging of the skin.

Reference is made herein to the binding of galectin-3 by MPC of theinvention. Binding occurs between the two molecules in in vitro assaysand there is a natural chemical bond possible between the reactivemoieties of the two compounds. In vitro assays rarely reflect, at amicroscopic and chemical level, the actual mechanisms that occur invivo. Binding of galectin-3 by MCP in vivo is an event that may bereversible to some degree—in the same fashion that an antibody may bindand then release a poorly held antigen. Galectin-3 is ubiquitous inmammals, found in many different tissues and spaces. Administration ofMCP does not appear to result in removal of all galectin-3 from thebody. By the term binding, Applicant intends to describe the formationof a complex between galectin-3 and MCP that removes the galectin-3 fromactive involvement in the metabolic and biological processes of thebody. Whether that complex is permanent and eliminated from the bodyover time, or reversible, does not appear to impact the relief obtained.

It is to be understood that MCP rarely mediates only inhibition ofcancer progression, fibrosis or inflammation. These are conditions thatare closely related, and tumor growth is frequently combined withinflammation and development of fibroses. Inflammation enhances thespeed with which a cancer progresses, so treatment of one may includetreatment of the other. Certainly, non-cancerous diseases, particularlyheart disease and atherosclerosis, arthritis and diabetes are associatedwith inflammation and fibrosis. There will be patients in need oftreatment primarily for inhibition of inflammation, patients in need oftreatment primarily for suppression of fibrosis, and patients whoseprimary indication is the progression or transformation of cancer. Whilepatient selection is a critical step in the methods embraced by thisinvention, it does not alter the fact that the patients receiving MCP inamounts of 5-1,500 mg/kg/day will receive the full benefit of MCP,independent of their primary indication.

The MCP of this invention may be the MCP of 10,000-20,000 Daltons, witha degree of etherification of less than about ten percent (10%). Furtherresearch has demonstrated that even lower molecular weight MCP may bemore efficacious, identifying a preferred range of 3,000-13,000 Daltonsmolecular weight. With this newer formulation has come a small revisionin the preferred range of administration. While 5-1,500 mg/kg/dayremains an effective range, more effective MCP may be administered in amore preferred range of 10 mg/kg/day to 750 mg/kg/day. The MCPs of thisinvention may be administered over a prolonged period of time, as manydisease conditions associated with inflammation and fibroticcomplications are chronic in nature. Extended administration of MCP hasnot shown toxicity or presented issues of tolerance in any way. Nocytopathic or toxicologic complications are associated with theadministration of MCP in this range either orally or intravenously.These MCP are readily absorbed intrabucally or through the intestinalmucosa into the bloodstream. In one alternative embodiment, the MCP maybe encapsulated, as a powder, into gelatin capsules which may, forexample, include 500-800 mg/capsule. Alternatively, a water-basedpreparation may be used, for example, six (6) capsules taken three (3)times a day with a full eight (8) ounces of water or juice. Controlleddosage formulations are preferred to ensure adequate, constant dosageover time.

MCPs may be combined with a wide variety of pharmaceutically acceptablecarriers, conventional excipients, flavorings and the like that aresuitable for oral or intravenous administration, depending on theprotocol desired. The modified pectins may also be administered togetherwith agents that will enhance complexation with galectin-3, such asglutathione-rich why protein and other binding adjuvants. Chelatingagents, such as 2,3 dimercapto-1-propane sulphonic acid andDL-2,3-dimercapto-succinic acid, EDTA may also be useful. MCP can beadded to formulations that include pharmaceuticals, botanicals, mineraland vitamins, to create additional effects, as needed. Absorption may beenhanced by using intrabuccal and transdermal delivery systems.

It should be noted that commonly, inflammation and fibrosis can beinduced by human agency, not just trauma or disease condition. Theadministration of MCP and its binding of galectin-3 can be effective inreducing or preventing organ damage induced by chemotherapy and otherpharmaceuticals. Some examples include bleomycin, which induces lungfibroses, and a wide variety of cardiac drugs such as amiodarone.Adriamycin and doxorubicin are widely prescribed and present cardiacinflammation and fibroses issues. Bacillus Calmette-Guerin washes totreat bladder cancer induce systemic inflammation and cyclophosphamidealso induces bladder damage. Ciclosporine, a widely usedimmunosuppressant drug, and the active agent in Restasis™, induceskidney toxicity and inflammation. Studies indicate that the vast arrayof organ damage caused by prescribed pharmaceuticals is mediated, atleast in part, by elevated galectin-3 levels, and can be limited if noteliminated by administration of MCP.

Examples Renal Injury

Renal insult is simulated in mice with folic acid. Folic acid inducedrenal injury candidates were pretreated with either water, or watersupplemented with one percent (1%) PectaSol-C one (1) week beforeinjection of folic acid. The gross changes associated with the renalinsult, including enlarged kidneys and weight loss, were reduced in micereceiving MCP. In the recovery phase, MCP-receiving mice demonstratedreduced galectin-3 and decreased renal fibrosis, macrophages,pro-inflammatory cytokine expression and apoptosis. The levels of otherrenal-associated galectins, including galectin-1 and galectin-9, wereunchanged. Clearly, MCP was of value in suppressing both inflammationand fibrosis relative to organ injury. Kolatsi-Joannou, PLoS One,8,6(4), e18683 (2011).

Thyroid Cancer Treatment

Patients with papillary thyroid cancer were administered a galectin-3binding molecule, inhibiting galectin-3 concentrations in the patientsso studied. Patients with reduced galectin-3 concentrations exhibitedimproved apoptotic activity, and improved sensitivity to both radiationand chemotherapeutic treatment (doxorubicin). Galectin-3 inhibition byadministration of a molecule that binds to galectin-3 offers a promisingtherapeutic treatment to both inhibit cancer, and elevate the utility ofconventional antineoplastic agents and treatments. Mol. Cancer Res.,7,10, 1655-62 (2009).

Lung Cancer Treatment

In patients with pulmonary adenocarcinoma, the test group wasadministered polyclonal antibodies which bind galectin-3 in much thesame fashion as MCP does. The tissues of these patients exhibited asignificant inhibition of cancer cell growth, the galectin-3 clearlyplaying a role in oncogenesis. Binding of galectin-3, as a means ofreducing its effective local concentration, whether by antibody or MCPoffers a therapeutic target for cancer intervention. Li et al, Clin.Invest. Med., 33, 1, e44-53 (2010).

Asthma and Related Inflammation

Mice with reduced galectin-3 concentrations were compared with mice withnormal galectin-3 levels in mice with induced asthma (ovalbumininduced). Mice with lower levels of galectin-3 exhibited fewereosinophils and lower goblet metaplasia, less airwayhyperressponsiveness and a different Th1:Th2 response. Administration ofMCP is an effective means of reducing the concentration of galectin-3 towhich cells, organs and systems sensitive to inflammation are exposed.This reduction provides benefits in treatment to intractable diseasestates. Zuberi et al, Am J. Pathol. 165, 6, 2045-53 (2004).

Inflammation of the Gastrointestinal Tract

Inflammation is a normal mammalian response to cellular stress in a widevariety of environments. In gastric ulcers, inflammation can oftenrepresent an imbalance in mucosal defense. Wistar albino rats withinduced gastric ulcers with a 3-fold reduction in galectin-3concentration exhibited improved protection against inflammation andgastric wall damage. Galectin-3 is, generally, an inflammationmodulator, and wherever that inflammation is a response to a conditionthat is injurious or imbalanced, MCP may be administered to reducegalectin-3 concentration locally and inhibit associated inflammation.Srikanta et al, Biochimie, 92, 2, 194-203 (2010).

Inflammation and Fibrosis of the Liver

Normal mice and galectin-3 deficient mice were compared after being feda diet that results in the formation of advanced lipoxidationendproducts or ALEs associated with inflammation and fibrosis.Galectin-3 deficient mice exhibited significantly reduced hepaticinflammation and fibrosis together with reduced hepatocyte injury.Reduced concentration of galectin-3 may also lead to reduced insulinresistance. MCP reduces effective concentration of galectin-3 in tissuesthreatened by inflammation and fibrosis. Reducing galectin-3concentration may be an effective therapeutic measure in addressingliver disease. Federici. J. Hepatol. 54,5,975-83 (2011).

Treatment of Liver Cancer

Balb/C mice were divided into a control group, and test groups thatreceived varying levels of MCP following administration of colon cancercells to the spleen to set up a colon cancer liver metastasis model. TheMCP was delivered in varying levels through the drinking water. Theconcentration of serum galectin-3 was significantly higher in thecontrol group. Expression of galectin-3 was found to significantlyincrease liver metastases of colon cancer. The administration of MCP toreduce the concentration of galectin-3 resulted in a significantreduction in liver metastases that varied directly with theconcentration of MCP in the drinking water (the higher theconcentration, the more dramatic the reduction in liver metastases.) Liuet al, World J. Gastroenterol. 14, 48, 7386-91 (2008).

Inflammation to Due Parasite Invasion

In mammals infected with Trypanasoma brucei, chronic inflammation is akey factor in the development of ACD. In mice with a reduced galectin-3concentration, significantly lower levels of anemia during infectionwere observed, and the mice survived twice as long as untreated mice.The mice with a reduced galectin-3 concentration reflected reduced liverpathology as well. Reduced inflammation was accompanied by reducedanemia and better survival, indicating reduction of galectin-3 levels isa potential therapeutic avenue for liver malfunction. MCP may beadministered to reduce effective galectin-3 levels. Vankrunkelsven,Immunobiology, 215, 9-10, 833-841 (2010). In a related study, galectin-3deficient mice demonstrated lower bacterial count when challenged with asublethal dose of Rhodococcus equi, together with a decreased frequencyof bacterial replication and survival. Ferraz et al, Eur. J Inmunol. 38,10, 2762-75 (2008). Reduced levels of galectin-3 in mice were alsoassociated with reduced inflammation in mice infected with Schistosomamansoni. Breuilh et al, Infect. Immun. 75, 11, 5148-57 (2007).

Diabetes Resistance

Mice were exposed to conditions that induce hyperglycemia and similardiabetic traits. In mice with reduced galectin-3 levels, measurements ofglycemia, quantitative histology and insulin content showed these miceto be resistant to the development of diabetes, as compared with micewith normal levels of galectin-3. The same mice showed a reduction ininflammation. One method of reducing the galectin-3 levels to whichchallenged tissues and organs are exposed is by systemic administrationof MCP, orally or intravenously. Reduction in galectin-3 levels isassociated with resistance to diabetogenesis. Mensah-Brown, Ann N.Y.Acad. Sci. 1084, 49-57 (2006). Related research has demonstrated thatreduction in galectin-3 levels slows the breakdown of the innerblood-retinal barrier (iBRB) that typically occurs early in diabetes.Galectin-3 deficient mice demonstrated a significant reduction indiabetes-mediated iBRB and reduced junctional disruption when comparedwith mice with normal galectin-3 levels. Canning et al, Exp. DiabetesRes., 2007:51837 (2007). Among the methods available to effectivelyreduce active galectin-3 concentrations to inhibit the development andprogression of diabetes and its symptoms is the administration of lowmolecular weight MCP over a long term. No toxicity has been demonstratedfor such administration.

Arthritis and Inflammation

A model of arthritis may be induced in mice by immunization withmethylated bovine serum albumin. Referred to as AIA, this conditionmimics arthritis and the inflammation associated with it. Inflammationwas shown to be markedly reduced, together with a reduction in boneerosion, in mice with reduced galectin-3 levels. The reduction inarthritis was accompanied by decreased levels of proinflammatorycytokines. Confirming that the nature or the galectin-3 level alterationcan be genetic or chemical, exogenously added glaectin-3 restored thelevel of arthritis in galectin-3 deficient mice to wild-type levels.Forsman et al, Arthritis Rheum., 63, 2, 445-54 (2011). Reduction ingalectin-3 levels as a means of addressing arthritis and relatedinflammation was also shown in rats where an artificial reduction ingalectin-3 levels via genetic modification substantially suppressedarthritis indices. Wang et al, Gene Ther., 17, 10, 1225-33 (2010).Administering low molecular weight MCP provides an effective in vivomethod of achieving this reduction of inflammation and treatingarthritis, including autoimmune arthritis such as rheumatoid arthritis.

Skin Inflammation

The development of inflammation in connection with allergic responsespresents a vast panorama of patient discomfort. A reduction ingalectin-3 levels in galectin-3 deficient mice was shown to reduceepidermal thickening, lower eosinophil infiltration and significantlyreduced dermatitis. Saegusa et al., Am. J. Pathol., 174, 3, 922-31(2009). Inflammation in a wide variety of tissues, as described above,is mediated by galectin-3, at least in part. Reducing the level ofactive galectin-3 by administration of MCP of molecular weight below20,000 Daltons down to 10,000 Daltons or lower, and preferably about3-13,000 Daltons, provides an effective, easily tolerated method ofreducing galectin-3 levels to achieve this goal. MCP can also be appliedtrans-dermally for such purposes.

Cardiac Disease and Fibrosis

As noted, MCP mediated reduction of galectin-3 levels may provide animportant treatment for cardiac diseases, particularly by reducingcardiac fibrosis. Reducing galectin-3 levels in the myocardium inosteopontin-deficient mice resulted in diminished fibrotic response andinflammation. Psarras et al, Eur. Heart. J., April 2011. Galectin-3levels associated with mediated fibrosis are much higher thanpost-fibrotic recovery values. De Boer et al, Eur. J. Heart Fail., 11,9, 811-17 (2009) suggesting that lowering galactin-3 levels temporarilyfollowing heart insult may reduce or suppress fibrosis and heart diseaseand failure associated therewith.

What has been clearly demonstrated is that reducing galectins-3 levelsin serum, cell and particularly in circulation may beneficially affecttissues and organs in mammals. It may also confer protection inmammalian patients presented with challenge from cancer cells, or cellsthat may develop into cancerous cells. Galectin-3 is a powerful mediatorof the development of inflammation and fibrosis and diseases andconditions associated therewith in a wide variety of tissue types, fromcardiac to kidney to liver, to lung, to skin. Further, administration ofMCP to a mammal in amounts of from 5 mg/kg/day on up to 1,500 mg/kg/davmay be effective in reducing and controlling inflammation throughout thebody.

While the present invention has been disclosed both generically and withreference to specific alternatives, those alternatives are not intendedto be limiting unless reflected in the claims set forth below. Theinvention is limited only by the provisions of those claims, and theirequivalents, as would be recognized by one of skill in the art to whichthis application is directed, in general, a medical doctor of at leastfive (5) years experience.

1-6. (canceled)
 7. A method of treating a mammal which benefits from areduction in available circulating galectin-3, comprising the steps of:a) Identifying a mammal in need of treatment for kidney damage, and b)Administering to said mammal an amount of modified pectin of molecularweight of 3,000-13,000 Daltons, in an amount of 10-750 mg/kg/day, for aperiod of time sufficient such that said mammal exhibits a reduction inactive galectin-3 levels in said mammal and thereby treat kidney damagein said mammal.
 8. A method of treating a mammal which benefits from areduction in available galectin-3, comprising the steps of: a)Identifying a mammal in need of treatment for kidney damage, and b)Administering to said mammal modified pectin of low molecular weight of10,000-20,000 Daltons, in an amount of 5-1,500 mg/kg/day, for a periodof time sufficient for said mammal to exhibit a reduction in activegalectin-3 levels in said mammal and thereby treat kidney damage in saidmammal.